In September 2019, a 22-year-old man with a history of drug abuse presented to the hospital with altered mental status. Due to a suspected drug overdose, a blood sample taken on admission and a urine sample collected 30 h thereafter were submitted to our laboratory to test for illegal drugs, pharmaceutical substances, and designer drugs. During the routine toxicological analysis of the serum sample, morphine and phenobarbital were identified by liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS). Additionally, two compounds showing identical accurate masses and isotope ratios as the designer benzodiazepine diclazepam and the benzodiazepine lormetazepam were found. However, retention times differed significantly from the expected values, and the acquired MS/MS spectra did not match the library entries of the two compounds, indicating the presence of two previously unknown substances. After further investigation, SL-164 (5-chloro-3-(4-chloro-2-methylphenyl)-2-methyl-4(3H)-quinazolinone), a methaqualone analog, which has recently emerged on the research chemical market, and its hydroxy metabolite were tentatively identified by accurate mass, isotope matching, and plausible fragmentation. However, for unequivocal confirmation and quantification, a reference standard is required. As no reference material was available by the end of 2019, SL-164 was obtained from an online shop, and its identity and purity (97.8%) were confirmed by nuclear magnetic resonance spectroscopy. The subsequent quantitative analysis revealed a concentration of 390 ng/mL SL-164 in serum. In the urine sample, the parent compound was not detected, but three suspected monohydroxylated metabolites were found. This example shows that LC-QTOF-MS is a powerful approach for the (tentative) identification of unknown compounds in biological matrices.

Synthetic cannabinoid receptor agonists (SCRAs) continue to show high prevalence on the new psychoactive substances drug market. Around 2019-2020, new SCRAs bearing a cumyl moiety emerged: Cumyl-CBMEGACLONE and Cumyl-NBMEGACLONE, carrying a cyclobutyl methyl (CBM) and a norbornyl methyl moiety (NBM) attached to the γ-carbolinone core. These were followed by Cumyl-NBMINACA, the indazole carboxamide analog of Cumyl-NBMEGACLONE. The study aimed at evaluating the human phase-I metabolism of these compounds and at identifying suitable urinary markers to prove their consumption. After enzymatic hydrolysis, 14 authentic urine samples (eight for Cumyl-CBMEGACLONE, four for Cumyl-NBMEGACLONE, and two for Cumyl-NBMINACA) were analyzed by liquid chromatography-quadrupole time-of-flight mass spectrometry. Results were compared with in vitro metabolites generated by pooled human liver microsomes incubation. Fifteen human phase-I metabolites were identified for Cumyl-CBMEGACLONE, nine for Cumyl-NBMEGACLONE, and thirteen for Cumyl-NBMINACA. The main in vivo metabolites were built by monohydroxylation, dihydroxylation, or trihydroxylation. The following urinary biomarkers are suggested for detecting the consumption of the investigated SCRAs: products of monohydroxylation at the CBM and at the core for Cumyl-CBMEGACLONE; two products of monohydroxylation at the norbonyl methyl tail for Cumyl-NBMEGACLONE; and metabolites built by dihydroxylation at the NBM substructure and by an additional hydroxylation at the cumyl moiety for Cumyl-NBMINACA.

This study investigates the presence of new psychoactive substances (NPS) and their metabolites in two wastewater treatment plants (WWTPs) situated in South Wales, UK (WWTP-1 and WWTP-2). Analysis was conducted for 35 NPS and metabolites, along with the inclusion of benzoylecgonine (main cocaine metabolite) and cannabis, the most detected illicit substances. Benzoylecgonine was identified as the predominant substance in both WWTPs. Epidemiological calculations revealed the average population consumption of cocaine to be 3.88 mg/d/1000 inhabitants around WWTP-1 and 1.97 mg/d/1000 inhabitants for WWTP-2. The removal efficiency of benzoylecgonine across both WWTPs was observed at an average of 73%. Subsequent qualitative analyses on randomly selected wastewater samples detected medicinal compounds including buprenorphine, methadone, and codeine in both WWTPs. An additional experiment employing enzymatic hydrolysis revealed the presence of morphine, an increased presence of codeine, and 11-Nor-9-Carboxy-THC (THC-COOH) post-hydrolysis. These findings underscore the significant presence of illicit substances and medicinal compounds in wastewater systems with the absence of NPS within the South Wales area, highlighting the necessity for enhanced monitoring and treatment strategies to address public health and environmental concerns.

This six-year multicentre study investigated acute intentional poisoning with substances of abuse in adolescents to identify changes and patterns in substance use. Data from 562 adolescents were collected from three paediatric poison centres in Romania between January 2017 and December 2022. This study analysed the epidemiological and sociodemographic characteristics of the adolescents, including age, gender, place of residence, history of substance abuse, psychiatric history, and history of institutionalised care. The findings revealed that cannabis and new psychoactive substances (NPSs) are the most commonly implicated substances, each with distinct profiles among adolescents. Cannabis was involved in 46.1% of cases, with a significant association with urban residency. NPSs were identified as the second most prevalent substance, accounting for 39.3% of cases. These were more prevalent in rural areas and among patients with psychiatric disorders. Cannabis and NPSs were also the most commonly implicated substances in acute intentional poisoning cases with substances of abuse. These substances have distinct profiles among adolescents, including age, gender, residency area, history of substance abuse, psychiatric history, and institutional care. These findings underscore the necessity of targeted public health interventions and integrated care approaches to address substance use and related mental health issues in adolescents.

Hundreds of new psychoactive substances (NPS) covering most drugs-of-abuse classes have been introduced to the recreational drug market in recent years. One class of NPS drugs that has become more common recently is \"designer\" benzodiazepines. Due to a close structural resemblance with prescription benzodiazepines, some of these substances may elicit a positive response (i.e. cross react) in immunoassay screening. Consequently, it is increasingly important to include NPS benzodiazepines during method confirmation to ensure accurate identification of closely-related compounds as well as detection of the benzodiazepines themselves. Here, we present our efforts to develop a screening and confirmation method for detection of 28 NPS benzodiazepines in urine using reversed-phase liquid chromatographic separation in combination with high-resolution mass spectrometry (LC-HRMS). MS was performed in positive electrospray mode on a Thermo Fischer Scientific Q Exactive Orbitrap instrument using either full scan (for screening) or parallel reaction monitoring (for confirmation). We found the lower quantification limit of the method to range from 5 to 50 ng/mL. Analytical precision and accuracy were ≤15% for both screening and confirmation for all except one analyte. The method was used to analyze patient urine samples from routine drug testing and samples from acute intoxication cases presenting in emergency wards. Altogether, 16 of the 28 benzodiazepines (i.e., clobazam, clonazolam, deschloroetizolam, diclazepam, estazolam, etizolam, flubromazepam, flubromazolam, flunitrazolam, 3-hydroxyflubromazepam, 3-hydroxyphenazepam, ketazolam, meclonazepam, metizolam, nifoxipam, and pyrazolam) were detected in the urine samples. The results from patient sample analysis indicate a high prevalence of NPS benzodiazepine use, emphasizing the importance of including novel drugs of abuse in drug testing menus.

Novel benzodiazepine (NBz) detections in Victorian coronial cases started early in 2018 and have continued to increase in number and type up to December 2022. The eleven different NBz detections included etizolam (n=82), flualprazolam (n=43), clonazolam or 8-aminoclonazolam (n=30), bromazolam (n=15), clobromazolam (n=13), phenazepam (n=13), flubromazolam (n=12), flubromazepam (n=8), desalkylflurazepam (n=6), diclazepam (n=2), and estazolam (n=1). The pattern of detections varied over the 5-year period, with different compounds appearing over different time frames. The most recent NBz to appear were bromazolam, clobromazolam, flubromazepam and phenazepam; whereas etizolam had been seen regularly in case work since 2018. Of the total 133 deaths, 95 were considered drug related deaths by forensic pathologists with at least one additional CNS depressant also present capable of contributing to death. All deaths involved other (non-benzodiazepine) CNS active drugs, although many involved multiple NBz, with five or more different benzodiazepines detected in eight cases.

The complexity of the drug market and the constant updating of drugs have been challenging issues for drug regulatory authorities. With the emergence of new psychoactive substances (NPS) and the nonmedical use of prescription drugs, forensic and toxicology laboratories have had to adopt new drug screening methods and advanced instrumentation. Using high-performance liquid chromatography coupled with Orbitrap mass spectrometry, we developed a screening method for common NPS and other drugs. Two milliliters of mixed solvent of n-hexane and ethyl acetate (1:1, v:v) were added to 500 μL of blood or urine sample for liquid-liquid extraction, and methanol extraction was used for hair samples. The developed method was applied to 3897 samples (including 332 blood samples, 885 urine samples, and 2680 hair samples) taken from drug addicts in a province of China during 2019-2021. For urine and blood samples, the limits of detection (LODs) ranged from 1.68 pg/mL to 10.7 ng/mL. For hair samples, the LODs ranged from 3.30 × 10-5 to 4.21 × 10-3 ng/mg. The matrix effects of urine, blood, and hair samples were in the range of 47.6%-121%, 39.8%-139%, and 6.35%-118%, respectively. And the intra-day precision was 3.5%-6.0% and the inter-day precision was 4.18%-9.90%. Analysis of the actual samples showed an overall positive detection rate of 58.9%, with 5.32% of the samples indicating the use of multiple drugs.

BACKGROUND: In addition to the drugs that have been known for decades, several hundred mainly synthetic substances have been identified as drugs for the first time in the last 20 years.
OBJECTIVE: Presentation of the various groups of substances and their psychotropic effects, the epidemiology of their use and the legal and social background of this development.
METHODS: Narrative literature review.
RESULTS: The most important new psychoactive substances (NPS) are synthetic cannabinoids, synthetic stimulants (cathinones), halluginogens and new synthetic opioids (NSO), in particular fentanyl and related substances. The new substances do not have any qualitatively new psychotropic effects. They were brought onto the market in particular as substitutes for substances subject to the Narcotics Act but are often associated with dangerous side effects and even mortality. The increasing availability of these substances has gone hand in hand with the establishment of the Internet as a source of knowledge (e.g. for synthesis routes) and as a marketplace. Substance group-related regulations have also been established in Germany (New Psychoactive Substances Act). In Germany the prevalence of NPS use is significantly lower than that of cannabis; however, there are indications that the production and distribution of synthetic drugs is more profitable for drug dealers than with conventional plant-based drugs, such as heroin. In the USA, for example, NSOs are the primarily drugs used for opioid addiction.
CONCLUSIONS: It remains to be seen whether NPS and NSOs will replace conventional drugs. The availability of synthetic drugs is more difficult to reduce than that of plant-based drugs. Harm reduction measures should be expanded, e.g., early warning systems for new drugs, drug checking and naloxone programs.
UNASSIGNED: HINTERGRUND: Zusätzlich zu den seit Jahrzehnten bekannten Drogen sind in den letzten 20 Jahren mehrere hundert synthetische Substanzen als Drogen erstmals festgestellt worden.
UNASSIGNED: Darstellung der verschiedenen Stoffgruppen und ihrer psychotropen Wirkungen, der Epidemiologie ihres Konsums sowie der rechtlichen und sozialen Hintergründe dieser Entwicklung.
METHODS: Narrative Literaturübersicht.
UNASSIGNED: Die wichtigsten neuen psychoaktiven Substanzen (NPS) betreffen synthetische Cannabinoide, Stimulanzien und Halluzinogene sowie neue synthetische Opioide (NSO), insbesondere Fentanyl und verwandte Substanzen. Die neuen Substanzen weisen keine qualitativ neuen psychotropen Wirkungen auf. Sie wurden unter anderem als Ersatzstoffe für dem Betäubungsmittelgesetz unterliegende Substanzen auf den Markt gebracht, sind aber oft mit gefährlicheren Nebenwirkungen bis hin zu Mortalität belastet. Die steigende Verfügbarkeit dieser Substanzen ging parallel zur Etablierung des Internets als Wissensquelle (z. B. für Synthesewege) wie als Handelsplatz. Stoffgruppenbezogene Regularien wurden auch in Deutschland etabliert (Neue-Psychoaktive-Stoffe-Gesetz). In Deutschland ist die Prävalenz des Konsums von NPS z. B. im Vergleich zu Cannabis deutlich niedriger. Es gibt allerdings Hinweise, dass Produktion und Vertrieb synthetischer Drogen für Drogenhändler profitabler sind als Produktion und Vertrieb herkömmlicher pflanzlich basierter Drogen wie Heroin. In den USA sind NSO die vorrangig konsumierten Drogen bei Opioidabhängigkeit.
CONCLUSIONS: Es bleibt abzuwarten, ob NPS und NSO die herkömmlichen Drogen verdrängen. Die Verfügbarkeit synthetischer Drogen ist schwieriger zu reduzieren als die pflanzlich basierter Drogen. Maßnahmen zur Schadensminderung sollten ausgebaut werden, z. B. Frühwarnsysteme für neue Drogen, Drug Checking sowie Naloxon-Programme.

The production and use of New Psychoactive Substances (NPS) has skyrocketed over the last decade, causing major challenges for government authorities, public health agencies, and laboratories across the world. NPS are designed to mimic the psychoactive effects of unregulated or controlled drugs, while constantly being modified to evade drug control regulation. Hence, they are referred to as \"legal highs\", as they are technically legal to sell, possess, and use. NPS can be classified by their pharmacological mechanism of action and include cannabimimetic, depressants, dissociatives, hallucinogens, opioids, and stimulants. There is significant structural diversity within each NPS class, leading to variable detection using traditional clinical laboratory testing and complicating the interpretation of results. In this article, we review each of the NPS classes and summarize their associated mechanism of action, common structures, and metabolic pathways, and provide examples of recent drugs and emerging threats with a focus on Canadian drug trends. We also explore the current analytical advantages and limitations commonly faced by the clinical laboratory and provide insight on how toxicosurveillance can improve detection of NPS in the ever-changing NPS landscape.

Piperazines are a class of new psychoactive substances with hallucinogenic effects that affect the central nervous system by affecting the level of monoamine neurotransmitters. Abuse of piperazines will produce stimulating and hallucinogenic effects, accompanied by headache, dizziness, anxiety, insomnia, vomiting, chest pain, tachycardia, hypertension and other adverse reactions, and may even cause cardiovascular diseases and multiple organ failure and lead to death, seriously affecting human physical and mental health and public safety. The abuse of new psychoactive substance piperazines has attracted extensive attention from the international community. The study of its pharmacological toxicology and analytical methods has become a research hotspot in the field of forensic medicine. This paper reviews the in vivo processes, sample treatment and analytical methods of existing piperazines, in order to provide reference for forensic identification.
哌嗪类新精神活性物质是一类具有致幻作用的化合物，通过影响单胺能神经递质的水平进而影响中枢神经系统。滥用该类物质后，会产生刺激和致幻作用并伴头痛、头晕、焦虑、失眠、呕吐、胸痛、心动过速、高血压等不良反应，甚至可能造成心血管系统损害和多器官衰竭而死亡，严重危害身心健康和公共安全。哌嗪类新精神活性物质的滥用已引起国际社会的广泛关注，对其药理毒理及分析方法的研究成为法医学领域的研究热点。本文综述了已有的哌嗪类新精神活性物质的体内过程、样品处理以及分析方法，以期为法医学鉴定提供参考依据。.